Surface Roughness and Microhardness of Two Recent CAD/CAM-Materials After Storage

Objectives: To investigate surface roughness and microhardness of two recent resin-ceramic materials for computer-aided design/computer-aided manufacturing (CAD/CAM) after polishing with three polishing systems. Surface roughness and microhardness were measured immediately after polishing and after six months storage including monthly artificial toothbrushing. Methods: Sixty specimens of Lava Ultimate (3M ESPE) and 60 specimens of VITA ENAMIC (VITA Zahnfabrik) were roughened in a standardized manner and polished with one of three polishing systems (n=20/group): Sof-Lex XT discs (SOFLEX; three-step (medium-superfine); 3M ESPE), VITA Polishing Set Clinical (VITA; two-step; VITA Zahnfabrik), or KENDA Unicus (KENDA; one-step; KENDA Dental). Surface roughness (Ra; μm) was measured with a profilometer and microhardness (Vickers; VHN) with a surface hardness indentation device. Ra and VHN were measured immediately after polishing and after six months storage (tap water, 37°C) including monthly artificial toothbrushing (500 cycles/month, toothpaste RDA ~70). Ra- and VHN-values were analysed with nonparametric ANOVA followed by Wilcoxon rank sum tests (α=0.05). Results: For Lava Ultimate, Ra (mean [standard deviation] before/after storage) remained the same when polished with SOFLEX (0.18 [0.09]/0.19 [0.10]; p=0.18), increased significantly with VITA (1.10 [0.44]/1.27 [0.39]; p=0.0001), and decreased significantly with KENDA (0.35 [0.07]/0.33 [0.08]; p=0.03). VHN (mean [standard deviation] before/after storage) decreased significantly regardless of polishing system (SOFLEX: 134.1 [5.6]/116.4 [3.6], VITA: 138.2 [10.5]/115.4 [5.9], KENDA: 135.1 [6.2]/116.7 [6.3]; all p<0.0001). For VITA ENAMIC, Ra (mean [standard deviation] before/after storage) increased significantly when polished with SOFLEX (0.37 [0.18]/0.41 [0.14]; p=0.01) and remained the same with VITA (1.32 [0.37]/1.31 [0.40]; p=0.58) and with KENDA (0.81 [0.35]/0.78 [0.32]; p=0.21). VHN (mean [standard deviation] before/after storage) remained the same regardless of polishing system (SOFLEX: 284.9 [24.6]/282.4 [31.8], VITA: 284.6 [28.5]/276.4 [25.8], KENDA: 292.6 [26.9]/282.9 [24.3]; p=0.42-1.00). Conclusion: Surface roughness and microhardness of Lava Ultimate was more affected by storage and artificial toothbrushing than was VITA ENAMIC.

Dentin Bond Strength of Two Recent CAD/CAM-Materials after Storage

Purpose: To investigate the bond strength to dentin of two recent resin-ceramic materials for computer-aided design/computer-aided manufacturing (CAD/CAM) after 24 hours and after six months storage. Methods and Materials: Ninety cylinders were milled out of Lava Ultimate (3M ESPE) and 90 cylinders out of VITA ENAMIC (VITA Zahnfabrik) (dimension of cylinders: ∅=3.6 mm, h=2 mm). All Lava Ultimate cylinders were sandblasted (aluminium oxide, grain size: 27 μm) and cleaned with ethanol, whereas all VITA ENAMIC cylinders were acid-etched (5% hydrofluoric acid) and cleaned with water-spray. According to the three groups of cements used, the cylinders (n=30/resin-ceramic material) were further pretreated with 1) Scotchbond Universal for RelyX Ultimate (3M ESPE), 2) CLEARFIL Ceramic Primer for PANAVIA F2.0 (Kuraray), or 3) no further pretreatment for Ketac Cem Plus (3M ESPE). The cylinders were then bonded to ground human dentin specimens with 1) Scotchbond Universal and RelyX Ultimate (light-cured), 2) ED PRIMER II and PANAVIA F2.0 (light-cured), or 3) no adhesive system; Ketac Cem Plus (self-cured). Shear bond strength (SBS) was measured after 24 hours for 15 specimens/group and after six months (37°C, 100% humidity) for the other 15 specimens/group. SBS-values were statistically analysed with nonparametric ANOVA followed by exact Wilcoxon rank sum tests (α=0.05). Results: SBS of the two resin-ceramic materials and the three cements after 24 hours and after six months storage are shown in Figure 1. The statistical analysis showed that the duration of storage had a significant effect on SBS of Lava Ultimate for all three cements but had no significant effect on SBS of VITA ENAMIC. For Lava Ultimate SBS-values were (MPa; medians after 24 hours/six months): 13.5/22.5 (p=0.04) for RelyX Ultimate, 11.4/5.8 (p=0.0006) for PANAVIA F2.0, and 0.34/0.09 (p=0.04) for Ketac Cem Plus (Fig. 1). For VITA ENAMIC SBS-values were (MPa; medians after 24 hours/six months): 16.0/21.2 (p=0.10) for RelyX Ultimate, 11.4/14.4 (p=0.06) for PANAVIA F2.0, and 0.43/0.41 (p=0.32) for Ketac Cem Plus (Fig. 1). After 24 hours, there was no significant difference in SBS between Lava Ultimate and VITA ENAMIC for all three cements (p≥0.37). After six months, there was no significant difference in SBS between Lava Ultimate and VITA ENAMIC for RelyX Ultimate and Ketac Cem Plus (p≥0.07) whereas for PANAVIA F2.0, SBS was significantly lower for Lava Ultimate than for VITA ENAMIC (p<0.0001). Conclusion: SBS of Lava Ultimate was more affected by six months storage and by the cement used than was VITA ENAMIC.

Avian area vasculosa and CAM as rapid in vivo pro-angiogenic and antiangiogenic models.

Angiogenesis, the development of new blood vessels from preexisting ones, is driven by coordinated signaling pathways governed by specific molecules, hemodynamic forces, and endothelial and periendothelial cells. The processes involve adhesion, migration, and survival machinery within the target endothelial and periendothelial cells. Factors that interfere with any of these processes may therefore influence angiogenesis either positively (pro-angiogenesis) or negatively (antiangiogenesis). The avian area vasculosa (AV) and the avian chorioallantoic membrane (CAM) are two useful tools for studying both angiogenesis and antiangiogenesis since they are amenable to both intravascular and topical administration of target, agents, are relatively rapid assays, and can be adapted very easily to study angiogenesis-dependent processes, such as tumor growth. Both models provide a physiological setting that permits investigation of pro-angiogenic and antiangiogenic agent interactions in vivo.

Twelve percent of hips with a primary cam deformity exhibit a slip-like morphology resembling sequelae of slipped capital femoral epiphysis

BACKGROUND In some hips with cam-type femoroacetabular impingement (FAI), we observed a morphology resembling a more subtle form of slipped capital femoral epiphysis (SCFE). Theoretically, the morphology in these hips should differ from hips with a primary cam-type deformity. QUESTIONS/PURPOSES We asked if (1) head-neck offset; (2) epiphyseal angle; and (3) tilt angle differ among hips with a slip-like morphology, idiopathic cam, hips after in situ pinning of SCFE, and normal hips; and (4) what is the prevalence of a slip-like morphology among cam-type hips? METHODS We retrospectively compared the three-dimensional anatomy of hips with a slip-like morphology (29 hips), in situ pinning for SCFE (eight hips), idiopathic cam deformity (171 hips), and 30 normal hips using radial MRI arthrography. Normal hips were derived from 17 asymptomatic volunteers. All other hips were recruited from a series of 277 hips (243 patients) seen at a specialized academic hip center between 2006 and 2010. Forty-one hips with isolated pincer deformity were excluded. Thirty-six of 236 hips had a known cause of cam impingement (secondary cam), including eight hips after in situ pinning of SCFE (postslip group). The 200 hips with a primary cam were separated in hips with a slip-like morphology (combination of positive fovea sign [if the neck axis did not intersect with the fovea capitis] and a tilt angle [between the neck axis and perpendicular to the basis of the epiphysis] exceeding 4°) and hips with an idiopathic cam. We evaluated offset ratio, epiphyseal angle (angle between the neck axis and line connecting the center of the femoral head and the point where the physis meets the articular surface), and tilt angle circumferentially around the femoral head-neck axis. Prevalence of slip-like morphology was determined based on the total of 236 hips with cam deformities. RESULTS Offset ratio was decreased anterosuperiorly in idiopathic cam, slip-like, and postslip (eg, 1 o'clock position with a mean offset ranging from 0.00 to 0.14; p < 0.001 for all groups) compared with normal hips (0.25 ± 0.06 [95% confidence interval, 0.13-0.37]) and increased posteroinferiorly in slip-like (eg, 8 o'clock position, 0.5 ± 0.09 [0.32-0.68]; p < 0.001) and postslip groups (0.55 ± 0.12 [0.32-0.78]; p < 0.001) and did not differ in idiopathic cam (0.32 ± 0.09 [0.15-0.49]; p = 0.323) compared with normal (0.31 ± 0.07 [0.18-0.44]) groups. Epiphyseal angle was increased anterosuperiorly in the slip-like (eg, 1 o'clock position, 70° ± 9° [51°-88°]; p < 0.001) and postslip groups (75° ± 13° [49°-100°]; p = 0.008) and decreased in idiopathic cam (50° ± 8° [35°-65°]; p < 0.001) compared with normal hips (58° ± 8° [43°-74°]). Posteroinferiorly, epiphyseal angle was decreased in slip-like (eg, 8 o'clock position, 54° ± 10° [34°-74°]; p < 0.001) and postslip (44° ± 11° [23°-65°]; p < 0.001) groups and did not differ in idiopathic cam (76° ± 8° [61°-91°]; p = 0.099) compared with normal (73° ± 7° [59°-88°]) groups. Tilt angle increased in slip-like (eg, 2/8 o'clock position, 14° ± 8° [-1° to 30°]; p < 0.001) and postslip hips (29° ± 10° [9°-48°]; p < 0.001) and decreased in hips with idiopathic cam (-7° ± 5° [-17° to 4°]; p < 0.001) compared with normal (-1° ± 5° [-10° to 8°]) hips. The prevalence of a slip-like morphology was 12%. CONCLUSIONS The slip-like morphology is the second most frequent pathomorphology in hips with primary cam deformity. MRI arthrography of the hip allows identifying a slip-like morphology, which resembles hips after in situ pinning of SCFE and distinctly differs from hips with idiopathic cam. These results support previous studies reporting that SCFE might be a risk factor for cam-type FAI.

T1ρ MRI detects cartilage damage in asymptomatic individuals with a cam deformity

Hips with a cam deformity are at risk for early cartilage degeneration, mainly in the anterolateral region of the joint. T1ρ MRI is a described technique for assessment of proteoglycan content in hyaline cartilage and subsequently early cartilage damage. In this study, 1.5 Tesla T1ρ MRI was performed on 20 asymptomatic hips with a cam deformity and compared to 16 healthy control hips. Cam deformity was defined as an alpha angle at 1:30 o'clock position over 60° and/or at 3:00 o'clock position over 50.5°. Hip cartilage was segmented and divided into four regions of interest (ROIs): anterolateral, anteromedial, posterolateral and posteromedial quadrants. Mean T1ρ value of the entire weight bearing cartilage in hips with a cam deformity (34.0 ± 4.6 ms) was significantly higher compared to control hips (31.3 ± 3.2 ms, p = 0.050). This difference reached significance in the anterolateral (p = 0.042) and posteromedial quadrants (p = 0.041). No significant correlation between the alpha angle and T1ρ values was detected. The results indicate cartilage damage occurs in hips with a cam deformity before symptoms occur. A significant difference in T1ρ values was found in the anterolateral quadrant, the area of direct engagement of the deformity, and in the posteromedial quadrant. To conclude, T1ρ MRI can detect early chondral damage in asymptomatic hips with a cam deformity. This article is protected by copyright. All rights reserved.

CAD/CAM fabrication accuracy of long- vs. short-span implant-supported FDPs

OBJECTIVE To compare the precision of fit of long-span vs. short-span implant-supported screw-retained fixed dental prostheses (FDPs) made from computer-aided-design/computer-aided-manufactured (CAD/CAM) titanium and veneered with ceramic. The null hypothesis was that there is no difference in the vertical microgap between long-span and short-span FDPs. MATERIALS AND METHODS CAD/CAM titanium frameworks for an implant-supported maxillary FDP on implants with a flat platform were fabricated on one single master cast. Group A consisted of six 10-unit FDPs connected to six implants (FDI positions 15, 13, 11, 21, 23, 25) and group B of six 5-unit FDPs (three implants, FDI positions 21, 23, 25). The CAD/CAM system from Biodenta Swiss AG (Berneck, Switzerland) was used for digitizing (laser scanner) the master cast and anatomical CAD of each framework separately. The frameworks were milled (CAM) from a titanium grade V monobloc and veneered with porcelain. Median vertical distance between implant and FDP platforms from the non-tightened implants (one-screw test on implant 25) was calculated from mesial, buccal, and distal scanning electron microscope measurements. RESULTS All measurements showed values <40 μm. Total median vertical microgaps were 23 μm (range 2-38 μm) for group A and 7 μm (4-24 μm) for group B. The difference between the groups was statistically significant at implant 21 (P = 0.002; 97.5% CI -27.3 to -4.9) and insignificant at implant 23 (P = 0.093; -3.9 to 1.0). CONCLUSIONS CAD/CAM fabrication including laboratory scanning and porcelain firing was highly precise and reproducible for all long- and short-span FDPs. While all FDPs showed clinically acceptable values, the short-span FDPs were statistically more precise at the 5-unit span distance.

Dentinhaftkraft zweier neuer CAD/CAM-Materialien in Abhängigkeit verschiedener adhäsiver Zementsysteme

Zielsetzung: Ziel der Studie war die Bestimmung der Dentinhaftkraft von zwei so-genannten Hybridmaterialien für computer-aided design/computer-aided manufacturing (CAD/CAM) Restaurationen unter Anwendung von fünf verschiedenen Zementen vor und nach sechsmonatiger Lagerung. Materialien und Methoden: Aus extrahierten menschlichen Molaren wurden 300 Dentinprobekörper hergestellt (n=15 pro Gruppe; 10 Gruppen (2 Hybridkeramiken, 5 Zemente) je nach 24 h/nach sechsmonatiger Lagerung). Aus Hybridkeramikblöcken von Lava Ultimate (3M ESPE) und VITA ENAMIC (VITA Zahnfabrik) wurden Zylinder hergestellt, welche standardisiert aufgeraut wurden. Anschliessend wurden die Hybrid-keramikzylinder mit einem der folgenden fünf Zemente auf die Dentinprobekörper zementiert: mit den Kompositzementen RelyX Ultimate (3M ESPE), PANAVIA F2.0 (Kuraray), Variolink II (Ivoclar Vivadent), els cem (Saremco Dental AG) oder als Negativkontrollgruppe mit dem kunststoffmodifizierten Glasionomerzement Ketac Cem Plus (3M ESPE). Die Dentinhaftkraft der Hybridkeramikzylinder wurde einerseits nach 24 h und andererseits nach sechsmonatiger Lagerung via Scherkrafttest bestimmt. Nach dem Scherkrafttest wurde das Bruchmuster unter einem Lichtmikroskop bei 40-facher Vergrösserung beurteilt. Die Dentinhaftkraftwerte wurden mittels nichtparametrischer ANOVA gefolgt von exakten Wilcoxon Rangsummen-Tests statistisch analysiert (α=0,05). Die Beurteilung des Bruchmusters wurde deskriptiv ausgewertet. Resultate: Für die Hybridkeramik Lava Ultimate und nach 24 h erzielten die Kompositzemente RelyX Ultimate und Variolink II die höchsten Dentinhaftkraftwerte. Die Dentinhaftkraftwerte von RelyX Ultimate und Variolink II unterschieden sich nicht signifikant. Die Dentinhaftkraftwerte von PANAVIA F2.0 unterschieden sich ebenfalls nicht signifikant von denjenigen von RelyX Ultimate, waren jedoch signifikant tiefer als diejenigen von Variolink II. Unter allen Kompositzementen erzielte els cem die tiefsten Dentinhaftkraftwerte. Nach sechsmonatiger Lagerung waren die Dentinhaftkraftwerte für RelyX Ultimate die höchsten, gefolgt von Variolink II, von els cem und anschliessend von PANAVIA F2.0, welcher nach sechsmonatiger Lagerung die tiefsten Dentinhaftkraftwerte der Kompositzemente zeigte. Der kunststoffmodifizierte Glasionomerzement Ketac Cem Plus zeigte sowohl nach 24 h als auch nach sechsmonatiger Lagerung die tiefsten Dentinhaftkraftwerte. Für VITA ENAMIC war die Reihenfolge der Zemente nach Dentinhaftkraft nach 24 h ähnlich wie diejenige nach sechsmonatiger Lagerung: Die Dentinhaftkraft war für RelyX Ultimate und Variolink II am höchsten, gefolgt von PANAVIA F2.0, von els cem und schlussendlich von Ketac Cem Plus mit den tiefsten Dentinhaftkraftwerten. Nach 24 h und für alle fünf Zemente unterschieden sich die Dentinhaftkraftwerte zwischen Lava Ultimate und VITA ENAMIC nicht signifikant. Nach sechsmonatiger Lagerung unterschieden sich die Dentinhaftkraftwerte zwischen Lava Ultimate und VITA ENAMIC ebenfalls nicht signifikant für RelyX Ultimate und els cem im Gegensatz zu den Dentinhaftkraftwerten von PANAVIA F2.0, Variolink II und Ketac Cem Plus, welche signifikant tiefer waren für Lava Ultimate als für VITA ENAMIC. Das häufigste Bruch-muster war für Lava Ultimate nach 24 h und für VITA ENAMIC sowohl nach 24 h als auch nach sechsmonatiger Lagerung adhäsiv zwischen Dentin und Zement. Nach sechs-monatiger Lagerung war für Lava Ultimate das häufigste Bruchmuster tendenziell gemischte Brüche. Schlussfolgerung: Basierend auf den Resultaten kann gesagt werden, dass für beide Hybridkeramiken sowohl RelyX Ultimate als auch Variolink II empfohlen werden können. PANAVIA F2.0 kann für VITA ENAMIC empfohlen werden, für Lava Ultimate allerdings weniger, da die Dentinhaftkraft nach sechsmonatiger Lagerung abnahm. Von einer konventionellen (allerdings nicht indizierten und in dieser Studie experimentellen) Zemen-tierung der beiden Hybridkeramiken mit dem kunststoffmodifizierten Glasionomerzement Ketac Cem Plus muss abgeraten werden.

Einfluss von Politursystemen auf Oberflächenrauigkeit und mechanische Eigenschaften von CAD/CAM-Materialien

Zielsetzung: Das Ziel dieser Studie war, den Einfluss von drei Politursystemen auf die Oberflächenrauigkeit von verschiedenen Materialien für computer-aided design/computer-aided manufacturing (CAD/CAM) Restaurationen mittels Profilometrie sowie die mikromechanischen Eigenschaften der Materialien mittels Mikrohärtemessgerät zu analysieren. Materialien und Methoden: Von dem CAD/CAM-Kompositmaterial Paradigm MZ100 (3M ESPE), der CAD/CAM-Feldspatkeramik VITABLOCS Mark II (VITA Zahnfabrik) und den CAD/CAM-Hybridmaterialien Lava Ultimate (3M ESPE), VITA ENAMIC (VITA Zahnfabrik) und AMBARINO High-Class (Creamed) wurden je 60 Prüfkörper zugeschnitten, gekennzeichnet und standardisiert aufgerauht. Die standardisierte Aufrauhung wurde mit Baseline-Rauigkeitsmessungen überprüft (Ra und Rz; µm). Die Prüfkörper wurden mit einem von drei Politursystemen poliert (n=20 pro CAD/CAM-Material): 1) Sof-Lex Scheiben (Disc-System, 3 Politurschritte: medium, fein und superfein; 3M ESPE), 2) VITA Polishing Set Clinical (Silikonpolitursystem, 2 Politurschritte: medium und fein; VITA Zahnfabrik) oder 3) KENDA Nobilis (Silikonpolierer, 1 Politurschritt (universal); KENDA Dental). Nach Politur der Prüfkörper wurden Ra und Rz sowie die mikromechanischen Eigenschaften Oberflächenhärte (VHN; Vickers Härte) und Elastizitätsmodul (EM; GPa) gemessen. In den darauf folgenden sechs Monaten wurden die Prüfkörper in Leitungswasser gelagert und insgesamt sechs Mal einem maschinellem Zahnbürsten zugeführt. Anschliessend wurden erneut Ra und Rz sowie VHN und EM gemessen. Ra-, Rz-, VHN- und EM-Werte wurden mittels nichtparametrischer ANOVA global analysiert und die p-Werte mittels Bonferroni-Holm Korrektur für multiples Testen korrigiert. Als post-hoc Tests wurden Kruskal-Wallis-Tests sowie exakte Wilcoxon Rangsummen-Tests verwendet und die p-Werte wurden nicht korrigiert. Das Signifikanzniveau wurde auf α=0,05 festgelegt. Resultate: Für alle drei CAD/CAM-Hybridmaterialien ergaben Sof-Lex Scheiben nach der Politur die tiefste Oberflächenrauigkeit (d. h. die tiefsten Ra- und Rz-Werte), gefolgt von KENDA Nobilis und von dem VITA Polishing Set Clinical. Bei dem CAD/CAM-Kompositmaterial sowie bei der CAD/CAM-Feldspatkeramik ergaben Sof-Lex Scheiben und KENDA Nobilis ähnliche Resultate, gefolgt von dem VITA Polishing Set Clinical. Bei einigen CAD/CAM-Materialien zeigten sich – zum Teil in Abhängigkeit des Politursystems – nach maschinellem Zahnbürsten und Lagerung signifikant höhere Ra- und Rz-Werte. Die CAD/CAM-Materialien zeigten unabhängig des Politursystems und der Lagerung signifikant verschiedene VHN- und EM-Werte. Bei einigen CAD/CAM-Materialien zeigten sich – zum Teil ebenfalls in Abhängigkeit des Politursystems – nach maschinellem Zahn-bürsten und Lagerung signifikant tiefere VHN- und EM-Werte. Schlussfolgerungen: Die Wahl des Politursystems beeinflusste die Oberflächenrauigkeit der CAD/CAM-Materialien markant, wobei Sof-Lex Scheiben insgesamt die besten Politurresultate zeigten, gefolgt von dem Silikonpolierer KENDA Nobilis. Von der Verwendung des Silikonpolitursystems VITA Polishing Set Clinical muss eher abgeraten werden. Das CAD/CAM-Kompositmaterial Paradigm MZ100 und die CAD/CAM-Hybridmaterialien Lava Ultimate und AMBARINO High-Class als weichere und elastischere Materialien liessen sich insgesamt besser polieren, waren aber bezüglich mechanischer Eigenschaften anfälliger auf Lagerung als die härtere CAD/CAM-Feldspatkeramik VITABLOCS Mark II und das CAD/CAM-Hybridmaterial VITA ENAMIC.

Intercellular adhesion and membrane polarity in rat hepatocytes: Localization of cell -CAM 105 and other domain-specific membrane proteins {\it in situ\/} and {\it in vitro\/} by electron microscopy

Cell-CAM 105 has been identified as a cell adhesion molecule (CAM) based on the ability of monospecific and monovalent anti-cell-CAM 105 antibodies to inhibit the reaggregation of rat hepatocytes. Although one would expect to find CAMs concentrated in the lateral membrane domain where adhesive interactions predominate, immunofluorescence analysis of rat liver frozen sections revealed that cell-CAM 105 was present exclusively in the bile canalicular (BC) domain of the hepatocyte. To more precisely define the in situ localization of cell-CAM 105, immunoperoxidase and electron microscopy were used to analyze intact and mechanically dissociated fixed liver tissue. Results indicate that although cell-CAM 105 is apparently restricted to the BC domain in situ, it can be detected in the pericanalicular region of the lateral membranes when accessibility to lateral membranes is provided by mechanical dissociation. In contrast, when hepatocytes were labeled following incubation in vitro under conditions used during adhesion assays, cell-CAM 105 had redistributed to all areas of the plasma membrane. Immunofluorescence analysis of primary hepatocyte cultures revealed that cell-CAM 105 and two other BC proteins were localized in discrete domains reminscent of BC while cell-CAM 105 was also present in regions of intercellular contact. These results indicate that the distribution of cell-CAM 105 under the experimental conditions used for cell adhesion assays differs from that in situ and raises the possibility that its adhesive function may be modulated by its cell surface distribution. The implications of these and other findings are discussed with regard to a model for BC formation.^ Analysis of molecular events involved in BC formation would be accelerated if an in vitro model system were available. Although BC formation in culture has previously been observed, repolarization of cell-CAM 105 and two other domain-specific membrane proteins was incomplete. Since DMSO had been used by Isom et al. to maintain liver-specific gene expression in vitro, the effect of this differentiation system on the polarity of these membrane proteins was examined. Based on findings presented here, DMSO apparently prolongs the expression and facilitates polarization of hepatocyte membrane proteins in vitro. ^

Inactivation and self -association of CaM kinase: Effects of ATP, substrate binding domains, and isozymic forms

Calcium/calmodulin-dependent protein kinase II (CaM kinase) is a multifunctional Ser/Thr protein kinase, that is highly enriched in brain and is involved in regulating many aspects of neuronal function. We observed that forebrain CaM kinase from crude homogenates, cytosolic fractions and purified preparations inactivates and translocates into the particulate fraction following autophosphorylation. Using purified forebrain CaM kinase as well as recombinant $\alpha$ isozyme, we determined that the formation of particulate enzyme was due to enzyme self-association. The conditions of autophosphorylation determine whether enzyme self-association and/or inactivation will occur. Self-association of CaM kinase is sensitive to pH, ATP concentration, and enzyme autophosphorylation. This process is prevented by saturating concentrations of ATP. However, in limiting ATP, pH is the dominant factor, and enzyme self-association occurs at pH values $\rm{<}7.0.$ Site-specific mutants were produced by substituting Ala for Thr286, Thr253, or Thr305,306 to determine whether these sites of autophosphorylation affect enzyme inactivation and self-association. The only mutation that influenced these processes was Ala286, which removed the protective effect afforded by autophosphorylation in saturating ATP. Enzyme inactivation occurs in the presence and absence of self-association and appears predominantly sensitive to nucleotide concentration, because saturating concentrations of $\rm Mg\sp{2+}/ADP$ or $\rm Mg\sp{2+}/ATP$ prevent this process. These data implicate the ATP binding pocket in both inactivation and self-association. We also observed that select peptide substrates and peptide inhibitors modeled after the autoregulatory domain of CaM kinase prevented these processes. The $\alpha$ and $\beta$ isozymes of CaM kinase were characterized independently, and were observed to exhibit differences in both enzyme inactivation and self-association. The $\beta$ isozyme was less sensitive to inactivation, and was never observed to self-associate. Biophysical characterization, and transmission electron microscopy coupled with image analysis indicated both isozymes were multimeric, however, the $\alpha$ and $\beta$ isozymes appeared structurally different. We hypothesize that the $\alpha$ subunit of CaM kinase plays both a structural and enzymatic role, and the $\beta$ subunit plays an enzymatic role. The ramifications for the functional differences observed for inactivation and self-association are discussed based on potential structural differences and autoregulation of the $\alpha$ and $\beta$ isozymes in both calcium-induced physiological and pathological processes. ^

Los operativos de la CAM (Comando Anticomunista Mendoza)

Durante el gobierno de Löpez Rega, los primeros operativos comenzaron en el año 1974 actuando agrupados bajo en nombre de Alianza Anticomunista Argentina. En Mendoza, durante el gobierno de Cafiero, actuaba la CAM (comando anticomunista Mendoza), los cuales tenían como misión detener a los grupos que se oponían al gobierno. En muchos lugares de Mendoza se colocaron bombas, se destruyeron teatros, clubes, casas y vidas.