Microencapsulated pancreatic islets: a pathologic study.

Dog pancreatic islets isolated by an enzymatic digestion method were encapsulated in an alginate-poly L-lysine-alginate membrane. These microencapsulated pancreatic islets were cultured in vitro to study their ability of insulin secretion. Portions of these in vitro-cultured microencapsulated pancreatic islets were taken out for a viability dye exclusion study as well as for pathologic studies to correlate them with insulin secretion ability. We found that there was a strong correlation between them. Good insulin-secreting microcapsules showed well-preserved cell membranes and beta-cell granules. An in vitro culture for one to two days in RPMI-1640 made the islets more stable, the cellular surface became smoother and the beta-granules were in better shape. The microencapsulated pancreatic islets were also injected into the peritoneum of streptozotocin-induced diabetic CDF1 mice. Blood glucose levels dropped and stayed low for up to 60 days. But, when non-encapsulated dog pancreatic islets were used, the blood glucose levels remained low for only about 14 days. A small portion of the injected microcapsules were washed out at specific times for pathologic study. Up to 28 days after injection, only a few of the injected microcapsules showed pericapsular cellular infiltrate. However, after 56 days, most of the microcapsules showed dense pericapsular cellular infiltrate. Immunohistochemical analysis of these infiltrates showed that the majority of cells were fibroblasts and macrophages. Most of the cells located in the inner portion of the infiltrate were fibroblasts, while the macrophages were located mainly on the outer portion. Both scanning and transmission electron microscopy showed that the surface of the microcapsule outer wall was much smoother than the inner wall. The size of the microcapsules was approximately 0.6-0.8 mm and the thickness of the wall measured around 10 nm. The smaller the microcapsule is, the less chance there is of rupture with release of the xenographic islets. Once the wall of the transplanted microcapsules was ruptured, the inner surface showed more increased inflammatory cell and fibroblast infiltration than the outer surface.     

Rapid determination of zygosity and common aneuploidies from amniotic fluid cells using quantitative fluorescent polymerase chain reaction following genetic amniocentesis in multiple pregnancies

Following second-trimester twin amniocentesis, we used quantitative fluorescent polymerase chain reaction (QF-PCR) assays and polymorphic small tandem repeats (STR) for rapid determination of zygosity and common aneuploidies from amniotic fluid (AF) cells in four pregnancies with like-sex twins, fused placentae and inconclusive chorionicity. The first and the second cases were suspected to have inadvertent sampling of the same amniotic cavity twice. The first case showed a dizygotic (DZ) pattern and repeat amniocentesis was thus avoided. The second case was monozygotic (MZ) and was complicated by discordant fetal growth and twin-twin transfusion syndrome. The third case was associated with a co-twin malformation, occipital encephalocele. DNA studies revealed MZ twinning with a discordant structural defect. The fourth case was associated with co-twin abnormalities of cystic hygroma and hydrops fetalis. DNA studies showed DZ twinning with discordant structural and chromosomal defects. The QF-PCR assay with STR has the advantages of rapid determination of zygosity and common aneuploidies in AF cells. This simple test appears to be useful in the instances of possible inadvertent puncture of the same amniotic cavity twice during amniocentesis and of discordant fetal structural and/or chromosomal abnormalities following genetic amniocentesis in multiple pregnancies with uncertain chorionicity.     

Cytokines and lipopolysaccharide induce nitric oxide synthase in cultured rat pulmonary artery smooth muscle.

In the current study, we describe cytokine and Escherichia coli lipopolysaccharide (LPS) induction of nitric oxide (NO) synthase mRNA levels in cultured smooth muscle from rat pulmonary artery (RPASM). Exposure of RPASM to interleukin-1 beta, interferon-gamma, or LPS alone did not significantly affect NO synthesis, as determined by nitrite concentrations in media. Exposure to tumor necrosis factor-alpha caused a modest (2x) increase in nitrite production. In contrast, exposure to a combination of the above three cytokines and LPS caused a large increase in NO synthesis. Exposure of RPASM to this combination caused an increase in mRNA levels of NO synthase (as described by Northern blot analysis with 32P-cDNA probe to an inducible form of NO synthase present in murine macrophages) that was apparent as early as 4 h. Expression of the induced gene product after exposure to the cytokine and LPS mixture was evident by significant increases in nitrite production at 12 h. Production of nitrite was completely abolished in the presence of NG-monomethyl-L-arginine (NMA), and this inhibition was reversible by the addition of excess L-arginine. NO synthase mRNA levels were not affected by NMA. The nitrite production induced by the combination of cytokines and LPS was abolished by pretreating cells with cycloheximide. These data indicate that a combination of cytokines and LPS affect expression of the gene for the inducible form of NO synthase in cultured RPASM.     

The skin,tongue, and brain as favorable organs for hog cholera diagnosis by immunofluorescence

Summary Hog cholera virus antigens were found densely distributed in skin and tongue of pigs experimentally infected with hog cholera virus. The finding described here warrants the usage of ear biopsies for hog cholera diagnosis on a herd basis.     

Preparation and characterization of polymides derived from bis(methoxycarbonyl)terephthaloyl chloride by interfacial condensation

A series of polyimides was synthesized by interfacial condensation of bis(methoxycarbonyl)terephthaloyl choride ( 1 ) with aromatic diamines and polycyclic aliphatic diamines. 1 (mixed isomers) was synthesized from pyromellitic dianhydride and contains about 60% para isomers and 40% meta isomer. The poly(amic ester)s 3 had inherent viscosities of 0.85–0.24 dL/g. All the poly(amic ester)s were soluble in NMP, DMAc, DMF, and DMSO, except for the polymer prepared from diamantane-1,6-diamine. Through thermal treatment the poly(amic-ester)s formed imide structures. All the polyimides and poly(amic ester)s were amorphous due to X-ray diffraction studies. The glass transition temperatures of these polyimides were in the range of 255–340°C, and the 5% weight loss temperatures were between 485 and 640 °C in nitrogen atmosphere. The polyimides ( 4c and 4d ) with polycyclic aliphatic diamines such as adamantane-1,3-diamine and diamantane-1,6-diamine had good thermal stability, with a temperature of 5% weight loss above 485°C. The polyimides 4c and 4d also had high glass transition temperature of 340°C and 300°C, respectively.     

Dynamic mechanical properties of dental amalgams

The dynamic mechanical properties of two high-copper amalgams (Tytin and Dispersalloy) and two traditional amalgams (Aristalloy and Aristalloy with Zn) were measured over a temperature range of 0-70 degrees C and at frequencies of 0.1, 1, and 10 Hz by means of a DuPont DMA. Values of storage modulus (E') for the amalgams were equivalent to the Young's modulus (E) measured from static mechanical test methods, with Dispersalloy demonstrating the highest moduli. Values of E' decreased with increased temperature. E' of traditional amalgams decreased more rapidly than did those of the Cu-rich amalgams. Values of loss modulus (E") for Tytin were smaller than those of Dispersalloy and the two types of Aristalloy. High values of E" for the traditional amalgams correspond to a greater viscous behavior. Marked differences between the magnitude of tan delta and its temperature coefficients for traditional and high-copper amalgams were observed, which is indicative of differences in visco-elastic behavior between these two amalgam systems.     

Alterations in the conjunctival bacterial flora following a single dose of azithromycin in a trachoma endemic area

BACKGROUND/AIMS: The World Health Organisation has recommended repeated mass treatment of children in trachoma endemic areas with oral azithromycin. While chlamydia, the causative agent of trachoma, remains universally sensitive to azithromycin, there is concern that large scale programmes may alter the bacterial flora and induce resistance in streptococcal species. In this study the effect of a single dose of azithromcyin on the prevalence, species distribution, and resistance of conjunctival bacterial flora was determined. METHODS: Baseline and 14 day follow up bacterial cultures were taken from the conjunctivae of 121 children who reside in a trachoma endemic area of Nepal. 91 children were treated with azithromycin at baseline and 31 children received deferred treatment at the 14 day follow up. RESULTS: Although the prevalence of bacterial pathogens decreased significantly with azithromycin treatment, a significant change in the distribution of specific bacterial pathogens could not be demonstrated. Streptococcal resistance to azithromycin was found significantly more frequently after treatment. No change in the prevalence, distribution, or resistance pattern was found in the untreated control group. CONCLUSION: Repeated mass treatment of trachoma endemic areas with oral azithromycin will have an effect on bacterial flora. However, further work needs to be done to determine if this will have any clinical relevance.