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In vitro cytotoxicity of dental casting alloys over 8 months.

Although in vitro cytotoxicity tests have been a valuable part of the biological testing of dental casting alloys, these tests are generally limited by their short-term nature ( < 168 h). The objective of the current study was to measure the in vitro cytotoxicity of representative types of dental casting alloys over a relatively long-term interval (months) and compare longer-term cytotoxicity with that seen initially. Polished casting alloy samples were exposed to cell-culture medium for 30-day intervals. During the last 3 days of each interval, the medium was changed to provide a 3-day extract which was then placed on fibroblast or macrophage cultures for 48 h. The mitochondrial activity of the cells was measured and compared to control cultures to assess the cytotoxic effect of the alloys. The cytotoxic effect was plotted versus time of medium exposure. The total time of exposure was 8 months. For most alloys, the mitochondrial response was constant over the 8 months, indicating that the cytotoxic effect of the alloys did not change significantly after extended exposure to the medium. A trend of improving biological response was suspect for a reduced-gold alloy, but this trend was not statistically significant. The mitochondrial activity of the macrophages was less sensitive to the alloy extracts than the activity of the fibroblasts, which was significantly suppressed for several alloy types. However, mitochondrial activity of the macrophages was significantly increased for several alloys in the early time intervals. The total noble metal content of the alloys was not necessarily predictive of the biological response. Thus, it appears that in this indirect test of in vitro cytotoxicity, the short-term cytotoxicity is predictive of the longer-term for many types of dental casting alloys.

Analysis of Variance↗

Role of polymeric Tamm-Horsfall protein in cast formation: oligosaccharide and tubular fluid ions.

BACKGROUND: In acute tubular necrosis (ATN), distal tubules are obstructed by casts formed by tubular debris, cells, and Tamm-Horsfall protein (THP). Since there are Arginine-Glycine-Aspartate (RGD) and Leucine-Aspartate-Valine (LDV) adhesive sequences in human THP, there may be direct integrin-mediated binding of tubular cells to THP. Alternatively, polymerization of THP may result in entrapment of the cells in its gel. METHODS: Adhesion of LLC-PK(1) cells to THP-coated wells was directly measured. THP concentrate was dissolved in solutions which mimic urine from ATN (ATN-S), distal convoluted tubule (DCT-S), collecting duct (CD-S), and monomeric buffer (M buffer). THP was also denatured by either boiling or N-glycanase digestion. Gel formation of THP was then measured. Inhibition of LLC-PK(1) cell adhesion to collagen type I was measured with each solution, as well as after the collagen was pretreated with either monomeric (mTHP) or polymeric (pTHP) THP. The effect of pTHP on the settling rate of LLC-PK(1) cells in suspension was also measured. RESULTS: LLC-PK(1) cells did not directly adhere to THP, a finding against integrin-mediated binding as a mechanism for in vivo tubular cell/THP cast formation. The high electrolyte concentration of ATN-S and CD-S, however, was associated with pTHP gel formation. Moreover, cells suspended in pTHP remained in suspension. In cell adhesion studies, mTHP attenuated cell adhesion by binding to the matrix, whereas pTHP attenuated cell adhesion by trapping cells in its gel. An active process was involved since both pTHP gel formation and attenuation of cell adhesion were abolished by boiling or oligosaccharide removal with N-glycanase digestion. CONCLUSIONS: With renal ischemia and proximal tubule cell shedding, ATN and collecting duct fluid composition enhance THP gel formation and thus favor tubular cast formation and obstruction. The present in vitro results indicate the importance of oligosaccharide residues in mediating the effect of the THP on gel formation and potential cast formation in ATN.

Animals↗

Quantitative comparison of caste differences in honeybee hemolymph.

The honeybee, Apis mellifera, is an invaluable partner in agriculture around the world both for its production of honey and, more importantly, for its role in pollination. Honeybees are largely unexplored at the molecular level despite a long and distinguished career as a model organism for understanding social behavior. Like other eusocial insects, honeybees can be divided into several castes: the queen (fertile female), workers (sterile females), and drones (males). Each caste has different energetic and metabolic requirements, and each differs in its susceptibility to pathogens, many of which have evolved to take advantage of the close social network inside a colony. Hemolymph, arthropods' equivalent to blood, distributes nutrients throughout the bee, and the immune components contained within it form one of the primary lines of defense against invading microorganisms. In this study we have applied qualitative and quantitative proteomics to gain a better understanding of honeybee hemolymph and how it varies among the castes and during development. We found large differences in hemolymph protein composition, especially between larval and adult stage bees and between male and female castes but even between adult workers and queens. We also provide experimental evidence for the expression of several unannotated honeybee genes and for the detection of biomarkers of a viral infection. Our data provide an initial molecular picture of honeybee hemolymph, to a greater depth than previous studies in other insects, and will pave the way for future biochemical studies of innate immunity in this animal.

Algorithms↗

Bronchial casts of human lungs using negative pressure injection.

Negative-pressure casting techniques have been used for obtaining silicone rubber casts of the avian respiratory system, which contains minute air capillaries, noncompressible hollow bones, and highly flexible air sacs. The possibilities of this technique for the study of human airway diseases, which present technical difficulties similar to those of avian lungs, are investigated here. Left lungs from patients with various obstructive lung diseases or with normal lungs were fixed at autopsy under 25 cm H2O airway pressure with 2.5% phosphate-buffered glutaraldehyde. Cannulated, isolated lobes were placed in a vacuum chamber, and Dow Corning 734 RTV silicone elastomer, diluted with 10% low-viscosity silicone oil, was introduced into the airways at--10 kPa. Following complete polymerization, the tissue was microdissected to reveal the lung cast in situ, and histological sections were obtained for correlative studies. The tissue was then macerated in 5.25% sodium hypochlorite. The total time from fixation to finished cast was 3-4 days. Linear shrinkage of the elastomer was less than 1% in glutaraldehyde or water and between 1 and 2% in bleach. The negative-pressure injection technique enabled complete and accurate filling of airways to the alveolar duct level at physiological pressures and provided good delineation of blind cavities such as obstructed airways or ecstatic mucous gland ducts. The technique proved useful for the study of obstructive lung disease and should also prove useful for modelling aerosol deposition in diseased lungs.

Bronchi↗

Variation in allele frequencies among caste groups of the Dhangars of Maharashtra, India: an analysis with Wright's Fst statistic.

Variations in allelle frequencies at 11 serological and biochemical loci among 21 endogamous Dhangar castes of Maharashtra, India, have been studied using Wright's FST statistic. Gene differentiation among these castes is only about 4 per cent at these polymorphic loci. The distributional form of FST is explicable by random genetic drift at this early stage of differentiation of these caste groups. The empirical relationship between the average allele frequencies and the ration of within to total gene diversity of the caste groups is also studied.

Alleles↗

Consanguineous marriages and marriage payment: a study among three south Indian caste groups.

The present study aims at understanding the interrelations between consanguineous marriages and marriage payment. The data are collected from three castes inhabiting two regions of Nellore district, Andhra Pradesh, South India. It is evident from the study that the dowry system is more prevalent among higher castes, while the bride wealth system is more common among the lower castes in the hierarchy. Further, it can be seen that the dowry system is more prevalent in developed regions, while bride-wealth is more common in backward regions. Marriage payment is found to be less common in close kin marriages than in unrelated marriages. Most of the uncle-niece marriages are without any marriage payment, in all the castes. However, most of the matrilateral cross-cousin and patrilateral cross-cousin marriages are also without any marriage payment in the Devanga.

Consanguinity↗

The impact of caste on the growth of male Sikhs in Punjab, India.

This study is based on a cross-sectional sample of 442 Sikh boys and young male adults who were born in and around the town of Phagwara, in the state of Punjab, India, and stayed there all their lives. The sample comprised Sikh boys at three crucial phases of growth, at 5-6, 10-11 and 15-16 years, and young adults around 18 years of age. The sample comprised three distinct caste groups, viz. Jats, Ramgarhias and Ravidassias, belonging respectively to upper, middle and lower socioeconomic strata of the Sikh community. Differences in stature and body weight are particularly marked around early adolescence, and there is some indication of caste differences reappearing in young adults. In the case of body mass index, however, the differences seem most marked in late adolescence. There is no clear directional pattern to the way skinfolds change, but inter-caste differences become more marked with age. There is a suggestion of continuing growth beyond 16 years of age, and indications that the well-off groups grow more, as compared to the poor groups, during this period. Comparisons of young adults with older groups of the same caste indicate an increase in body weight with age, but smaller stature in the older groups. There is thus evidence for a secular stature increase among present-day Punjabi Sikhs.

Adolescent↗

Chemical corrosion in cast iron in soil-water medium.

Grey cast iron metal strips were allowed to rust in varying compositions of soil-water media under the controlled environment. The process of corrosion was monitored by non-electrochemical method. Assessment of the extent of corrosion was carried out, both visually and by the method of weight loss coupons. It was found that a 80:20 weight:volume percent (w/v%) composition caused the most severe case of corrosion over a period of seven days. It was also observed that the corrosion in cast iron obeyed the relation, D = ktn [10]. The value of 'n' increased as corrosion became more severe. Gravimetric analysis and evidence from the Scanning Electron Microscopy (SEM) proved that gamma-FeOOH was one of the intermediates of corrosion in grey cast iron in soil-water media. An attempt has also been made to propose a mechanism for the corrosion in cast iron strips in soil-water media. It was found to be consistent with the one proposed by McEnaney and Smith [11].

Corrosion↗

The genetic heritage of the earliest settlers persists both in Indian tribal and caste populations.

Two tribal groups from southern India--the Chenchus and Koyas--were analyzed for variation in mitochondrial DNA (mtDNA), the Y chromosome, and one autosomal locus and were compared with six caste groups from different parts of India, as well as with western and central Asians. In mtDNA phylogenetic analyses, the Chenchus and Koyas coalesce at Indian-specific branches of haplogroups M and N that cover populations of different social rank from all over the subcontinent. Coalescence times suggest early late Pleistocene settlement of southern Asia and suggest that there has not been total replacement of these settlers by later migrations. H, L, and R2 are the major Indian Y-chromosomal haplogroups that occur both in castes and in tribal populations and are rarely found outside the subcontinent. Haplogroup R1a, previously associated with the putative Indo-Aryan invasion, was found at its highest frequency in Punjab but also at a relatively high frequency (26%) in the Chenchu tribe. This finding, together with the higher R1a-associated short tandem repeat diversity in India and Iran compared with Europe and central Asia, suggests that southern and western Asia might be the source of this haplogroup. Haplotype frequencies of the MX1 locus of chromosome 21 distinguish Koyas and Chenchus, along with Indian caste groups, from European and eastern Asian populations. Taken together, these results show that Indian tribal and caste populations derive largely from the same genetic heritage of Pleistocene southern and western Asians and have received limited gene flow from external regions since the Holocene. The phylogeography of the primal mtDNA and Y-chromosome founders suggests that these southern Asian Pleistocene coastal settlers from Africa would have provided the inocula for the subsequent differentiation of the distinctive eastern and western Eurasian gene pools.

Asia, Central↗

An improved technic for examining urinary casts and a review of their significance.

Casts represent an important, diagnostic component of urinary sediment, and may signal renal parenchymal disease in asymptomatic individuals. Accurate, precise identification of certain casts may be difficult due to poor visualization by current technics. Casts from freshly voided urine specimens, cytocentrifuged and stained by the Papanicolaou method, can be visualized optimally from permanent slide preparations. The genesis of these casts and their diagnostic potential related to renal disease and acute renal allograft rejection are discussed.

Erythrocytes↗

Delayed graft function and cast nephropathy associated with tacrolimus plus rapamycin use.

Delayed graft function (DGF) occurs in 15 to 25% (range, 10 to 62%) of cadaveric kidney transplant recipients and up to 9% of living donor recipients. In addition to donor, recipient, and procedural factors, the choice of immunosuppression may influence the development of DGF. The impact of immunosuppression on DGF was studied. The frequency of DGF was evaluated in first cadaveric or living donor kidney allograft recipients (n = 144) transplanted at the University of Washington from November 1999 through September 1, 2001. Donor, recipient, and procedural factors, as well as biopsy results, were compared between patients who developed DGF and those who did not. DGF was more common in patients treated with rapamycin than without (25% versus 8.9%, P = 0.02) and positively correlated with rapamycin dose (P = 0.008). In those developing DGF, the duration of posttransplant dialysis increased with donor age (P = 0.003) but decreased with mycophenolate mofetil use (P = 0.01). All biopsies during episodes of DGF demonstrated changes of acute tubular injury. Of the patients with tubular injury, 12 treated with rapamycin and tacrolimus developed intratubular cast formation indistinguishable from myeloma cast nephropathy. Histologic, immunohistochemical, and ultrastructural studies indicated that these casts were composed at least in part of degenerating renal tubular epithelial cells. These findings suggest that rapamycin therapy exerts increased toxicity on tubular epithelial cells and/or retards healing, leading to an increased incidence of DGF. Additionally, rapamycin treatment combined with a calcineurin inhibitor may lead to extensive tubular cell injury and death and a unique form of cast nephropathy.

Adult↗

Caste totipotency and conflict in a large-colony social insect.

In most social insects with large, complex colonies workers and queens are morphologically quite distinct. This means that caste determination must occur prior to adulthood. However workers and queens in the swarm-founding epiponine wasps are often morphologically indistinguishable, or nearly so, suggesting that caste determination in these wasps could be quite different. To determine the extent of caste lability in the epiponine, Parachartergus colobopterus, we removed all the queens from one colony and all but one from another colony. Worker aggression diminished after queen removal. A week later the colony with no queens had a new, young cohort of mated queens. These must have been either adults or pupae at the time of queen removal, and so could not have been fed any differently from workers. Relatedness patterns confirmed that these new queens would normally have been workers and not queens. A model of inclusive fitness interests shows that workers ought to suppress new queen production, except at low queen numbers, a prediction supported by our empirical results. The patterns of social conflict over queenship resulting from swarm founding in a many-queen society may help to explain the unusually weak caste differentiation in the epiponines.

Animals↗

Thickness dependence of structural relaxation in spin-cast, glassy polymer thin films.

The isothermal structural relaxation of glassy, spin-cast polymer thin films has been investigated. Specifically, the thickness h of freshly cast poly(methyl methacrylate) thin films was measured over time using spectroscopic ellipsometry. The spin-cast films exhibit a gradual decrease in thickness, which is attributed to structural relaxation of the glass combined with simultaneous solvent loss. In all cases, h was found to be greater than the equilibrium thickness h(infinity) , which is obtained by cooling slowly from the melt. It is observed that both the rate of the volume relaxation and the fractional departure from h(infinity) (referred to as delta(0) ) increase with increasing film thickness. In the limit of very thin films, the initial h is close to h(infinity) , and delta(0) is small, whereas in thick films (>500 nm) , a plateau value of delta(0) of 0.16 is observed, which is close to the volume fraction of the solvent at the vitrification point. This dependence of delta(0) on thickness is observed regardless of the substrate, polymer molecular weight, or angular velocity during spin casting. Enhanced mobility near film surfaces could be leading to greater relaxation in thinner films prior to, and immediately after, the vitrification of the polymer during the deposition process.

Journal Article↗

A system for digital reconstruction of gypsum dental casts.

A range scanner is developed that can scan a gypsum dental cast and reconstruct the cast digitally for display and storage purposes. The scanner is based on subtractive light and computes the range values using optical triangulation. A fiducial marker is introduced that, when attached to a dental cast at the time of image acquisition, makes it possible to integrate multiview range images of the cast without image registration. A method for calibrating the scanner is described and experimental results showing the accuracy of the scanner are presented.

Algorithms↗

Hair casts.

The literature on hair casts suggests they are rare even though Kligman intimated in 1957 that in some scalp diseases they might well be common. In this study, casts have been looked for in a group of orthokeratotic and parakeratotic scalp conditions. Hair casts are indeed common in psoriasis, pityriasis amiantacea, pityriasis capitis and seborrhoeic dermatitis. No casts were seen in women with traction hair styles or in those regularly using hair sprays.

Dermatitis, Seborrheic↗

ELKS, a protein structurally related to the active zone protein CAST, is involved in Ca2+-dependent exocytosis from PC12 cells.

The active zone protein CAST binds directly to the other active zone proteins RIM, Bassoon and Piccolo, and it has been suggested that these protein-protein interactions play an important role in neurotransmitter release. To further elucidate the molecular mechanism, we attempted to examine the function of CAST using PC12 cells as a model system. Although PC12 cells do not express CAST, they do express ELKS, a protein structurally related to CAST. Endogenous and exogenously expressed ELKS, RIM2 and Bassoon were colocalized in punctate signals in PC12 cells. Over-expression of full-length ELKS resulted in a significant increase in stimulated exocytosis of human growth hormone (hGH) from PC12 cells, similar to the effect of full-length RIM2. This increase was not observed following over-expression of deletion constructs of ELKS that lacked either the last three amino acids (IWA) required for binding to RIM2 or a central region necessary for binding to Bassoon. Moreover, over-expression of the NH(2)-terminal RIM2-binding domain of Munc13-1, which is known to inhibit the binding between RIM and Munc13-1, inhibited the stimulated increase in hGH secretion by full-length RIM2. Furthermore, this construct also inhibited the stimulated increase in hGH secretion induced by full-length ELKS. These results suggest that ELKS is involved in Ca(2+)-dependent exocytosis from PC12 cells at least partly via the RIM2-Munc13-1 pathway.

Adaptor Proteins, Signal Transducing↗

Caste-specific cytochrome P450 in the damp-wood termite Hodotermopsis sjostedti (Isoptera, Termopsidae).

Termites are eusocial insects with a well-defined caste system, which is an example of polyphenism. This polyphenism is based on hormonally controlled differential gene expression. In the damp-wood termite Hodotermopsis sjostedti, we induced differentiation into the soldier caste by using juvenile hormone analogue treatment. We then investigated specific gene expression, which appeared during the hormonal response and triggered caste differentiation, using fluorescent differential display. A candidate cDNA sequence with similarity to cytochromes P450, CYP6AM1, was characterized and its transcript shown to be repressed between 1 and 3 days after hormone treatment. CYP6AM1 was specifically expressed in the fat body of pseudergates and soldiers. The putative function of this P450 is discussed with respect to the caste differentiation system.

Amino Acid Sequence↗

Scanning electron microscopy of renal vascular casts.

The use of Cementex to form casts of the renal vascular system is described. These casts are useful in delineating the vascular microarchitecture of the kidney including that of the glomerular capillaries, but do not appear to reproducibly reflect physiological vascular tone. The postglomerular vascular system is often incompletely filled using this technique. The amount of filling can be altered by the techniques used in cast preparation. The effect of perfusion pressure, concentration of Cementex, fixation, and type of colour additive are compared with respect to the cast structure.

Animals↗